Document Detail


Application of semi-nested polymerase chain reaction targeting internal transcribed spacer region for rapid detection of panfungal genome directly from ocular specimens.
MedLine Citation:
PMID:  17595473     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: The incidence of fungal endophthalmitis has dramatically increased in recent years and rapid detection of fungi using nucleic acid-based amplification techniques is helpful in management. AIM: To evaluate semi-nested polymerase chain reaction (PCR) targeting internal transcribed spacer (ITS) region for detection of panfungal genome in ocular specimens. STATISTICAL ANALYSIS USED: Z test for two proportion. MATERIALS AND METHODS: Standardization of PCR targeting ITS primers was carried out by determining analytical sensitivity and specificity. The sensitivity and specificity of PCR was determined by serial tenfold dilutions of C. albicans (ATCC 24433) DNA and DNA extracts of laboratory isolates of Aspergillus fumigatus, Fusarium lichenicola (4), other fungal and closely related bacterial strains and also human DNA. Semi-nested PCR was applied onto a total of 168 ocular specimens with clinically suspected fungal etiology during 2003-2005. RESULTS AND CONCLUSIONS: PCR was specific and sensitive to detect 1fg of fungal DNA with ITS primers. PCR detected fungal genome in 90 (53.57%) in comparison with the conventional technique, positive in 34 (20.23%) by smear examination and in 42 (25%) by culture. The increase in clinical sensitivity by 28.57% using PCR was found to be statistically significant { P < 0.001 using Z test for two proportion}. The accuracy of the test was found to be 70.85%. PCR proved to be a rapid diagnostic technique for detection of panfungal genome directly from clinical specimens.
Authors:
R Bagyalakshmi; K L Therese; H N Madhavan
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Indian journal of ophthalmology     Volume:  55     ISSN:  0301-4738     ISO Abbreviation:  Indian J Ophthalmol     Publication Date:    2007 Jul-Aug
Date Detail:
Created Date:  2007-06-27     Completed Date:  2007-09-11     Revised Date:  2008-09-03    
Medline Journal Info:
Nlm Unique ID:  0405376     Medline TA:  Indian J Ophthalmol     Country:  India    
Other Details:
Languages:  eng     Pagination:  261-5     Citation Subset:  IM    
Affiliation:
Larsen and Toubro Microbiology Research Centre, Vision Research Foundation, No.18, College Road, Sankara Nethralaya, Chennai - 600 006, India.
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MeSH Terms
Descriptor/Qualifier:
Aqueous Humor / microbiology
Cornea / microbiology
DNA, Fungal / genetics*
Diagnosis, Differential
Endophthalmitis / diagnosis,  microbiology*
Eye Infections, Fungal / diagnosis,  microbiology*
Fungi / genetics*,  isolation & purification
Genome, Fungal / genetics*
Humans
Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Vitreous Body / microbiology
Chemical
Reg. No./Substance:
0/DNA, Fungal

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